Search results for: strain specific markers

Authors: Momita Das, Sofia Banu, Jibon Kotoky

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Environmental contamination of natural resources with persistent organic pollutants is of great world-wide apprehension. Polycyclic aromatic hydrocarbons (PAHs) are among the organic pollutants, released due to various anthropogenic activities. Due to their toxic, carcinogenic and mutagenic properties, PAHs are of environmental and human concern. Presently, bioremediation has evolved as the most promising biotechnology for cleanup of such contaminants because of its economical and less cost effectiveness. In the present study, distribution of 16 USEPA priority PAHs was determined in the soil samples collected from fifteen different sites of Guwahati City, the Gateway of the North East Region of India. The total concentrations of 16 PAHs (Σ16 PAHs) ranged from 42.7-742.3 µg/g. Higher concentration of total PAHs was found more in the Industrial areas compared to all the sites (742.3 µg/g and 628 µg/g). It is noted that among all the PAHs, Naphthalene, Acenaphthylene, Anthracene, Fluoranthene, Chrysene and Benzo(a)Pyrene were the most available and contain the higher concentration of all the PAHs. Since microbial activity has been deemed the most influential and significant cause of PAH removal; further, twenty-three bacteria were isolated from the most contaminated sites using the enrichment process. These strains were acclimatized to utilize naphthalene and anthracene, each at 100 µg/g concentration as sole carbon source. Among them, one Gram-positive strain (JMG-01) was selected, and biodegradation ability and initial catabolic genes of PAHs degradation were investigated. Based on 16S rDNA analysis, the isolate was identified as Bacillus cereus strain JMG-01. Topographic images obtained using Scanning Electron Microscope (SEM) and Atomic Force Microscope (AFM) at scheduled time intervals of 7, 14 and 21 days, determined the variation in cell morphology during the period of degradation. AFM and SEM micrograph of biomass showed high filamentous growth leading to aggregation of cells in the form of biofilm with reference to the incubation period. The percentage degradation analysis using gas chromatography and mass analyses (GC-MS) suggested that more than 95% of the PAHs degraded when the concentration was at 500 µg/g. Naphthalene, naphthalene-2-methy, benzaldehyde-4-propyl, 1, 2, benzene di-carboxylic acid and benzene acetic acid were the major metabolites produced after degradation. Moreover, PCR experiments with specific primers for catabolic genes, ndo B and Cat A suggested that JMG-01 possess genes for PAHs degradation. Thus, the study concludes that Bacillus cereus strain JMG-01 has efficient biodegrading ability and can trigger the clean-up of PAHs contaminated soil.

Keywords: AFM, Bacillus cereus strain JMG-01, degradation, polycyclic aromatic hydrocarbon, SEM

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Authors: Masoud Sakhinia, Sajjad Ahmad

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Introduction: Though knowledge of the compositional makeup and structure of the limbal niche has progressed exponentially during the past decade, much is yet to be understood. Identifying the precise profile and role of the stromal makeup which spans the ocular surface may inform researchers of the most optimum conditions needed to effectively expand LESCs in vitro, whilst preserving their differentiation status and phenotype. Limbal fibroblasts, as opposed to corneal fibroblasts are thought to form an important component of the microenvironment where LESCs reside. Methods: The corneal stroma was tissue engineered in vitro using both limbal and corneal fibroblasts embedded within a tissue engineered 3D collagen matrix. The effect of these two different fibroblasts on LESCs and hTCEpi corneal epithelial cell line were then subsequently determined using phase contrast microscopy, histolological analysis and PCR for specific stem cell markers. The study aimed to develop an in vitro model which could be used to determine whether limbal, as opposed to corneal fibroblasts, maintained the stem cell phenotype of LESCs and hTCEpi cell line. Results: Tissue culture analysis was inconclusive and required further quantitative analysis for remarks on cell proliferation within the varying stroma. Histological analysis of the tissue-engineered cornea showed a comparable structure to that of the human cornea, though with limited epithelial stratification. PCR results for epithelial cell markers of cells cultured on limbal fibroblasts showed reduced expression of CK3, a negative marker for LESC’s, whilst also exhibiting a relatively low expression level of P63, a marker for undifferentiated LESCs. Conclusion: We have shown the potential for the construction of a tissue engineered human cornea using a 3D collagen matrix and described some preliminary results in the analysis of the effects of varying stroma consisting of limbal and corneal fibroblasts, respectively, on the proliferation of stem cell phenotype of primary LESCs and hTCEpi corneal epithelial cells. Although no definitive marker exists to conclusively illustrate the presence of LESCs, the combination of positive and negative stem cell markers in our study were inconclusive. Though it is less traslational to the human corneal model, the use of conditioned medium from that of limbal and corneal fibroblasts may provide a more simple avenue. Moreover, combinations of extracellular matrices could be used as a surrogate in these culture models.

Keywords: cornea, Limbal Stem Cells, tissue engineering, PCR

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Authors: Suleyman Karabal, Timothy L. Burnett, Egemen Avcu, Andrew H. Sherry, Philip J. Withers

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SA508 Grade 3 steel is widely used in the construction of nuclear pressure vessels, where its fracture toughness plays a critical role in ensuring operational safety and reliability. Understanding the ductile failure mechanisms in this steel grade is crucial for designing robust pressure vessels that can withstand severe nuclear environment conditions. In the present study, round bar specimens of SA508 Grade 3 steel with four distinct notch geometries were subjected to tensile loading while capturing continuous 2D images at 5-second intervals in order to monitor any alterations in their geometries to construct true stress-strain curves of the specimens. 3D reconstructions of X-ray computed tomography (CT) images at high-resolution (a spatial resolution of 0.82 μm) allowed for a comprehensive assessment of the influences of second-phase particles (i.e., manganese sulfide inclusions and cementite particles) on ductile failure initiation as a function of applied plastic strain. Additionally, based on 2D and 3D images, plasticity modeling was executed, and the results were compared to experimental data. A specific ‘two-parameter criterion’ was established and calibrated based on the correlation between stress triaxiality and equivalent plastic strain at failure initiation. The proposed criterion demonstrated substantial agreement with the experimental results, thus enhancing our knowledge of ductile fracture behavior in this steel grade. The implementation of X-ray CT and fractography analysis provided new insights into the diverse roles played by different populations of second-phase particles in fracture initiation under varying stress triaxiality conditions.

Keywords: ductile fracture, two-parameter criterion, x-ray computed tomography, stress triaxiality

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Authors: Sourav Ghosh, Antony Gomes

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Since 1894 anti-snake venom serum (ASVS) is the only available treatment against snake envenomation, although there are many side effects and limitations. The need for a supportive treatment was felt for a long time to overcome the side effects and limitations of ASVS. Andrographolide conjugated with gold nanoparticle (A-GNP) has been found to antagonize viper venom-induced local damages. The present study was aimed to study the protective efficacy of A-GNP against Viper venom-induced inflammatory response and organ toxicity in animal model. Ethical clearance was obtained from animal experiments. Physico-chemical characterization of A-GNP was done by DLS (diameter and zeta potential), FE-SEM and XRD. Swiss albino male mice were divided into 4 groups: Gr.1-Sham control, Gr.2- Russell’s Viper venom (RVV) control, Gr.3- andrographolide treated and Gr.4- A-GNP treated. The 1/5th minimum lethal dose of RVV (500µg/kg, s.c.) was induced in animals of group 2, 3 & 4 animals, followed by treatment with andrographolide (100mg/kg, i.p.) and A-GNP (100mg/kg, i.v.) in group 3 & 4 animals, respectively. Blood was collected after 18 h, serum was prepared, and inflammatory markers (IL 1β, 6, 17a, 10, TNF α) and biochemical markers (AST, ACP, LDH, urea, creatinine) were assessed. Values were expressed as mean±SEM (n=4), one way ANOVA was done, P<0.05 was considered as statistically significant. DLS size showed the hydrodynamic diameter of A-GNP to be 230-260nm with polydispersity index of 0.103 and zeta potential was -18.32mV. XRD data confirmed the presence of crystalline gold in A-GNP, and FESEM indicated the presence of nearly spherical particle with size18-24nm.Treatment with A-GNP significantly decreased viper venom-induced proinflammatory markers (IL 1β, 6, 17, TNF α) increased anti-inflammatory markers (IL 10) and decreased organ toxicity markers (AST, ACP, LDH, urea, creatinine) in animal model. Venom neutralization efficacy of A-GNP was > andrographolide, which confirmed the increased efficacy of andrographolide after gold nanoparticle conjugation. Venom neutralization by A-GNP was due to anti-oxidant/anti-inflammatory activity of andrographolide, which showed increased efficacy after gold nanoparticle tagging. Thus, A-GNP may serve as a supportive therapy in snake-bite (against inflammatory response and organ toxicity) subject to further detail studies.

Keywords: andrographolide, gold nanoparticle, inflammatory response, organ toxicity, snake venom, snake venom neutralization, viper venom

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Authors: Mai A. Aljaberi

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The anchorage of fibre-reinforced polymer (FRP) sheets is the most effective solution to prevent or delay debonding failure; this system has proven to get better levels of FRP utilization. Unfortunately, the related design information is still unclear. This shortcoming limits the widespread use of the anchorage system. In order to minimize the knowledge gap about the design of U-wrap anchors, this paper reports the results of tested beams which were strengthened with carbon fiber-reinforced polymer (CFRP) sheets at their tension sides and secured with U-wrap anchors at each end of the longitudinal CFRP. The beams were tested under four-point loading until failure. The parameters examined include the compressive strength of the concrete and the number of longitudinal CFRP. It is concluded that these parameters have a considerable effect on the debonding of the strain. The greatest improvement in the strain was 55.8% over the control beam.

Keywords: CFRP, concrete strengthening, debonding failure, debonding strain, U-wrap anchor

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Authors: H. Gholami, M. Jalali Azizpour

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For materials such as hard coating whose stresses state are difficult to obtain by a widely used method called high-speed hole-drilling method (ASTM Standard E837). It is important to develop a non contact method. This process itself imposes an additional stresses. The through thickness residual stress of st37 steel using elector-discharge was investigated. The strain gage and dynamic strain indicator used in all cases was FRS-2-11 rosette type and TML 221, respectively. The average residual stress in depth of 320 µm was -6.47 MPa.

Keywords: HVOF, residual stress, thermal spray, WC-Co

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Authors: Martyna Chotomska, Aleksandra Studzinska, Marta Lisowska, Justyna Szubert, Aleksandra Tabis, Jacek Bania, Arkadiusz Miazek

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Objectives: Assessing antigen-specific T cell responses is of utmost importance for the pre-clinical testing of prototype vaccines against intracellular pathogens and tumor antigens. Mainly two types of in vitro assays are used for this purpose 1) enzyme-linked immunospot (ELISpot) and 2) intracellular cytokine staining (ICS). Both are time-consuming, relatively expensive, and require manual dexterity. Here, we assess if a straightforward detection of luciferase activity in blood samples of transgenic reporter mice expressing a secreted Lucia luciferase under the transcriptional control of IFN-γ promoter parallels the sensitivity of IFNγ ELISpot assay. Methods: IFN-γ-LUCIA mouse strain carrying multiple copies of Lucia luciferase transgene under the transcriptional control of IFNγ minimal promoter were generated by pronuclear injection of linear DNA. The specificity of transgene expression and mobilization was assessed in vitro using transgenic splenocytes exposed to various mitogens. The IFN-γ-LUCIA mice were immunized with 50mg of ovalbumin (OVA) emulsified in incomplete Freund’s adjuvant three times every two weeks by subcutaneous injections. Blood samples were collected before and five days after each immunization. Luciferase activity was assessed in blood serum. Peripheral blood mononuclear cells were separated and assessed for frequencies of OVA-specific IFNγ-secreting T cells. Results: We show that in vitro cultured splenocytes of IFN-γ-LUCIA mice respond by 2 and 3 fold increase in secreted luciferase activity to T cell mitogens concanavalin A and phorbol myristate acetate, respectively but fail to respond to B cell-stimulating E.coli lipopolysaccharide. Immunization of IFN-γ-LUCIA mice with OVA leads to over 4 fold increase in luciferase activity in blood serum five days post-immunization with a barely detectable increase in OVA-specific, IFNγ-secreting T cells by ELISpot. Second and third immunizations, further increase the luciferase activity and coincidently also increase the frequencies of OVA-specific T cells by ELISpot. Conclusions: We conclude that minimally invasive monitoring of luciferase secretions in blood serum of IFN-γ-LUCIA mice constitutes a sensitive method for evaluating primary and memory Th1 responses to protein antigens. As such, this method may complement existing methods for rapid immunogenicity assessment of prototype vaccines.

Keywords: ELISpot, immunogenicity, interferon-gamma, reporter mice, vaccines

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Authors: Sabitha Rani A., Nagamani V.

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Embelia ribes Burm.f (Family-Myrsinaceae) commonly known as Vidanga or Baibirang, is one of the important medicinal plants of India. The seed extract is reported to be antidiabetic, antitumour, analgesic, anti-inflammatory, antispermatogenic, free radical scavenging activities and widely used in more than 75 Ayurvedic commercial formulations. Among the 100 different species of Embelia, E. ribes is considered as a major source of Embelin, a bioactive compound. Because of high demand and low availability, the seeds of E. ribes are substituted with many cheaper alternatives. Therefore, the present study of RAPD-PCR analysis was undertaken to develop molecular markers for identification of E. ribes. A total of 13 different seed samples of Embelia were collected from different agro-climatic regions of India. The seeds of E.ribes were collected from Kalpetta, Kerala and three different seed samples were collected from traders of Odisha, Madhya Pradesh, Maharastra. The other nine seed samples were collected from local traders which they have collected from different regions of India. Genomic DNA was isolated from different seed samples E. ribes and RAPD-PCR was performed on 13 different seed samples using 47 random primers. Out of all the primers, only 22 primers produced clear and highly-reproducible banding patterns. The 22 selected RAPD primers generated a total of 280 alleles with an average of 12 alleles per primer pair. In the present study, we have identified three RAPD-PCR markers i.e. OPF5_480 bp, OPH11_520 bp and OPH4_530 bp which can be used for genetic fingerprinting of E. ribes. This methodology can be employed for identification of original E. ribes and also distinguishing it from other substitutes and adulterants.

Keywords: Embelia ribes, RAPD-PCR, primers, genetic analysis

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Authors: M. Mydlárová Blaščáková, J. Bernasovská, J. Poráčová, I. Boroňová

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Obesity is frequent attendant phenomenon of patients with endocrinological disease. Between BMI and endocrinological diseases is close correlation. In thesis we focused on the allocation of hormone concentration – PTH and TSH, CHOL a mineral element Ca in a blood serum. The examined group was formed by 100 respondents (women) aged 36 – 83 years, who were divided into two groups – control group (CG), group with diagnosed endocrine disease (DED). The concentration of PTH and TSH, Ca and CHOL was measured through the medium of analyzers Cobas e411 (Japan); Cobas Integra 400 (Switzerland). At individuals was measured body weight as well as stature and thereupon from those data we enumerated BMI. On the basis of Student T-test in biochemical parameter of PTH and Ca we found out significantly meaningful difference (p<0,05) between CG and DED. In CG we made a founding the association between BMI and PTH by means of correlation analysis.

Keywords: biochemical markers, hormones, obesity, women

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Authors: Madiha Z. Ammari, Abdul Qader AlNajmi

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A new system of U-links was used in this study to confine the concrete core in concrete-filled steel beams. This system aims to employ the separation expected between the steel tube and the concrete core in the compression side of the section in the plastic hinge zone. A total of six rectangular CFT beam specimens were tested under flexure using different D/t ratios and different diameters for the U-links to examine their effect on the flexural behavior of these beams. The ultimate flexural strength of the CFT beam specimens with U-links showed an increase of strength about 47% of the specimen with D/t ratio equals 37.5 above standard CFT beam specimen without U-links inside. State of concrete inside the tubes has shown no crushing of concrete when those beams were cut open at the location of the plastic hinge. Strain measurements revealed that the compressive strain of concrete was 5-6 times the concrete crushing strain.

Keywords: concrete-filled tubes, U-links, plated studies, beams, flexural strength, concrete, confinement

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Authors: Clement Ndoricimpa

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The expression of stance is highly expected in writing at tertiary level. Through a selection of linguistic and rhetorical elements, writers express commitment, critical distance and build a critically discerning reader in texts. Despite many studies on patterns of stance in students’ academic writing, little may not be known about how English as a Foreign Language students learns to build a critically discerning reader in their texts. Therefore, this study examines patterns of stance in essays written by students majoring in business administration at English-medium University in Burundi as part of classroom assignments. It draws on systemic functional linguistics to analyze qualitatively and quantitatively the data. The quantitative analysis is used to identify the differences in frequency of stance patterns in the essays. The results show a significant difference in the use of boosters by lower- and upper-level students. Lower-level students’ writing contains more boosters and many idiosyncratic sentence structures than do upper-level students’ writing, and upper-level students’ essays contain more hedging and few grammatical mistakes than do lower-level students’ essays. No significant difference in the use of attitude markers and concessive and contrastive expressions. Students in lower- and upper-level do not use attitude markers and disclaimer markers appropriately and accurately. These findings suggest that students should be taught the use of stance patterns in academic writing.

Keywords: academic writing, metadiscourse, stance, student corpora

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Authors: Mehdi Mohebodini, Iman Khalili-Baseri, Mehdi Behnamian, Sara Dezhsetan

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Diversity analysis at the molecular level using PCR-based markers is the efficient and rapid method of identifying the relationships and differences among the genotypes. In the present study, genetic diversity and relationships among 20 collected purslane accessions were evaluated using ISSR markers. The genotyping data were used to understand the relationships among the collected accessions and identify genetically diverse purslane accessions. The 25 primers gave a total of 92 bands, of which 62 were polymorphic (67.4%). The genetic diversity as estimated by Shannon’s information index was 0.55, revealing a quite high level of genetic diversity in the germplasm. The average number of an observed allele, effective allele, polymorphic information content (PIC) and Nei’s index were 2, 1.65, 0.37 and 0.37, respectively.

Keywords: Portulaca oleracea L., genetic diversity, ISSR, germplasm

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Authors: Dorcas Ropo Abejide, Olamide Ahmed Falusi, Oladipupo Abdulazeez Yusuf Daudu, Bolaji Zuluqurineen Salihu, Muhammad Liman Muhammad

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This study evaluated the morpho-agronomic response of twenty-four (24) Nigerian Bambara groundnut landraces to water stress and genetic diversity of some selected accessions using SSR markers. The studies were carried out in the botanical garden of the Department of Plant Biology, Federal University of Technology, Minna, Niger State, Nigeria in a randomized complete block design using three replicates. Molecular analysis using SSR primers was carried out at the International Institute of Tropical Agriculture (IITA) Ibadan in order to characterize ten selected accessions comprising the seven most drought tolerant and three most susceptible accessions from the 24 accessions evaluated. Results revealed that water stress decreased morpho-agronomic traits such as plant height, leaf area, number of leaves per plant, seed yield, etc. A total of 22 alleles were detected by the SSR markers used with a mean number of 4 allelles. SSR markers MBamCO₃₃, Primer 65, and G358B2-D15 each detected 4 allelles, while Primer 3FR and 4FR detected 5 allelles each. The study revealed significantly high polymorphisms in 10 Loci. The mean value of polymorpic information content was 0.6997, implying the usefulness of the primers used in identifying genetic similarities and differences among the Bambara groundnut genotypes. The SSR analysis revealed a comparable pattern between genetic diversity and drought tolerance of the genotypes. The UPGMA dendrogram showed that at a genetic distance of 0.1, the accessions were grouped into three groups according to their level of tolerance to drought. The two most drought-tolerant accessions were grouped together, and the 5th and 6th most drought-tolerant accessions were also grouped together. This suggests that the genotypes grouped together may be genetically close, may possess similar genes, or have a common origin. The degree of genetic variants obtained from this profiling could be useful in Bambara groundnut breeding for drought tolerance. The identified drought tolerant Bambara groundnut landraces are important genetic resources for drought stress tolerance breeding programme of Bambara groundnut. The genotypes are also useful for germplasm conservation and global implications.

Keywords: Bambara groundnut, genetic diversity, germplasm, SSR markers, water stress

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Authors: Vathna Suy, Ki-Il Song

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In order to store highly pressurized gas such as an isothermal compressed air energy storage, Lined Rock Caverns (LRC) are constructed underground and supported by layers of concrete, steel and rock masses. This study aims to numerically investigate the performance of rock masses which serve as a support of Lined Rock Cavern subjected to high cyclic pressure loadings. FLAC3D finite different software is used for the simulation since the software can effectively model the behavior of concrete lining and steel plate with its built-in structural elements. Cyclic pressure loadings are applied onto the inner surface of the cavern which then transmitted to concrete, steel and eventually to the surrounding rock masses. Changes of stress and strain are constantly monitored throughout all the process of loading operations. The results at various monitoring locations are then extracted and analyzed to assess the response of the rock masses, specifically on its ability to absorb energy during loadings induced by the changes of cyclic pressure loadings inside the cavern. By analyzing the obtained data of stress-strain relation and taking into account the behavior of materials under the effect of strain-dependency, conclusions on the performance of rock masses subjected to high cyclic loading conditions are drawn.

Keywords: cyclic loading, FLAC3D, lined rock cavern (LRC), strain-dependency

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Authors: Racholsan Raj Nirmal, B. S. V. Patnaik, R. Jayaganthan

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The deformation behaviour of additively manufactured AlSi10Mg alloy under low strains, high strain rates and elevated temperature conditions is essential to analyse and predict its response against dynamic loading such as impact and thermomechanical fatigue. The constitutive relation of Johnson-Cook is used to capture the strain rate sensitivity and thermal softening effect in AlSi10Mg alloy. Johnson-Cook failure model is widely used for exploring damage mechanics and predicting the fracture in many materials. In this present work, Johnson-Cook material and damage model parameters for additively manufactured AlSi10Mg alloy have been determined numerically from four types of uniaxial tensile test. Three different uniaxial tensile tests with dynamic strain rates (0.1, 1, 10, 50, and 100 s^-1) and elevated temperature tensile test with three different temperature conditions (450 K, 500 K and 550 K) were performed on 3D printed AlSi10Mg alloy in ABAQUS/Explicit. Hexahedral elements are used to discretize tensile specimens and fracture energy value of 43.6 kN/m was used for damage initiation. Levenberg Marquardt optimization method was used for the evaluation of Johnson-Cook model parameters. It was observed that additively manufactured AlSi10Mg alloy has shown relatively higher strain rate sensitivity and lower thermal stability as compared to the other Al alloys.

Keywords: ABAQUS, additive manufacturing, AlSi10Mg, Johnson-Cook model

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Authors: Syed Beenish Rufai, Sarman Singh

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Background: Performance of Genotype MTBDRsl (Hain Life science GmbH Germany) for detection of mutations associated with second-line drug resistance is well known. However, less evidence regarding the association of mutations and genetic background of strains is known which, in the future, is essential for clinical management of anti-tuberculosis drugs in those settings where the probability of particular genotype is predominant. Material and Methods: During this retrospective study, a total of 259 MDR-TB isolates obtained from pulmonary TB patients were tested for second-line drug susceptibility testing (DST) using Genotype MTBDRsl VER 1.0 and compared with BACTEC MGIT-960 as a reference standard. All isolates were further characterized using spoligotyping. The spoligo patterns obtained were compared and analyzed using SITVIT_WEB. Results: Of total 259 MDR-TB isolates which were screened for second-line DST by Genotype MTBDRsl, mutations were found to be associated with gyrA, rrs and emb genes in 82 (31.6%), 2 (0.8%) and 90 (34.7%) isolates respectively. 16 (6.1%) isolates detected mutations associated with both FQ as well as to AG/CP drugs (XDR-TB). No mutations were detected in 159 (61.4%) isolates for corresponding gyrA and rrs genes. Genotype MTBDRsl showed a concordance of 96.4% for detection of sensitive isolates in comparison with second-line DST by BACTEC MGIT-960 and 94.1%, 93.5%, 60.5% and 50% for detection of XDR-TB, FQ, EMB, and AMK/CAP respectively. D94G was the most prevalent mutation found among (38 (46.4%)) OFXR isolates (37 FQ mono-resistant and 1 XDR-TB) followed by A90V (23 (28.1%)) (17 FQ mono-resistant and 6 XDR-TB). Among AG/CP resistant isolates A1401G was the most frequent mutation observed among (11 (61.1%)) isolates (2 AG/CP mono-resistant isolates and 9 XDR-TB isolates) followed by WT+A1401G (6 (33.3%)) and G1484T (1 (5.5%)) respectively. On spoligotyping analysis, Beijing strain (46%) was found to be the most predominant strain among pre-XDR and XDR TB isolates followed by CAS (30%), X (6%), Unique (5%), EAI and T each of 4%, Manu (3%) and Ural (2%) respectively. Beijing strain was found to be strongly associated with D94G (47.3%) and A90V mutations by (47.3%) and 34.8% followed by CAS strain by (31.6%) and 30.4% respectively. However, among AG/CP resistant isolates, only Beijing strain was found to be strongly associated with A1401G and WT+A1401G mutations by 54.5% and 50% respectively. Conclusion: Beijing strain was found to be strongly associated with the most prevalent mutations among pre-XDR and XDR TB isolates. Acknowledgments: Study was supported with Grant by All India Institute of Medical Sciences, New Delhi reference No. P-2012/12452.

Keywords: tuberculosis, line probe assay, XDR TB, drug susceptibility

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Authors: Mayandi Ramanathan

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Gas turbine blades see the most aggressive thermal stress conditions within the engine, due to high Turbine Entry Temperatures in the range of 1500 to 1600°C. The blades rotate at very high rotation rates and remove a significant amount of thermal power from the gas stream. At high temperatures, the major component failure mechanism is a creep. During its service over time under high thermal loads, the blade will deform, lengthen and rupture. High strength and stiffness in the longitudinal direction up to elevated service temperatures are certainly the most needed properties of turbine blades and gas turbine components. The proposed advanced Ti alloy material needs a process that provides a strategic orientation of metallic ordering, uniformity in composition and high metallic strength. The chemical composition of the proposed Ti alloy material (25% Ta/(Al+Ta) ratio), unlike Ti-47Al-2Cr-2Nb, has less excess Al that could limit the service life of turbine blades. Properties and performance of Ti-47Al-2Cr-2Nb and Ti-6Al-4V materials will be compared with that of the proposed Ti alloy material to generalize the performance metrics of various gas turbine components. This paper will involve the summary of the effects of additive manufacturing and heat treatment process conditions on the changes in the phase composition, grain structure, lattice structure of the material, tensile strength, creep strain rate, thermal expansion coefficient and fracture toughness at different temperatures. Based on these results, additive manufacturing and heat treatment process conditions will be optimized to fabricate turbine blade with Ti-43Al matrix alloyed with an optimized amount of refractory Ta metal. Improvement in service temperature of the turbine blades and corrosion resistance dependence on the coercivity of the alloy material will be reported. A correlation of phase composition and creep strain rate will also be discussed.

Keywords: high temperature materials, aerospace, specific strength, creep strain, phase composition

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Authors: Kobid Panthi, Suttisak Soralump, Suriyon Prempramote

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In June 27, 2014 slope movement was observed in upstream side of Khlong Pa Bon Dam, Thailand. The slide did not have any major catastrophic impact on the dam structure but raised a very important question; why did the slide occur after 10 years of operation? Various site investigations (Bore Hole Test, SASW, Echo Sounding, and Geophysical Survey), laboratory analysis and numerical modelling using SIGMA/W and SLOPE/W were conducted to determine the cause of slope movement. It was observed that the dam had undergone the greatest differential drawdown in its operational history in the year 2014 and was termed as the major cause of movement. From the laboratory tests, it was found that the shear strength of clay had decreased with a period of time and was near its residual value. The cyclic movement of water, i.e., reservoir filling and emptying was coined out to be the major cause for the reduction of shear strength. The numerical analysis was carried out using a modified cam clay (MCC) model to determine the strain softening behavior of the clay. The strain accumulation was observed in the slope with each reservoir cycle triggering the slope failure in 2014. It can be inferred that if there was no major drawdown in 2014, the slope would not have failed but eventually would have failed after a long period of time. If there was no major drawdown in 2014, the slope would not have failed. However, even if there hadn’t been a drawdown, it would have failed eventually in the long run.

Keywords: slope movement, strain softening, residual strength, modified cam clay

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Authors: Dan Xie

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The use of discourse markers (DMs) can play a crucial role in representing discourse interaction and pragmatic competence. Learners’ use of DMs and differences between native speakers (NSs) and non-native speakers (NNSs) in the use of various DMs have been the focus of considerable research attention. However, some commonly used DMs, such as you know, have not received as much attention in comparative studies, especially in the Chinese context. This study analyses data in two corpora (COLSEC and Spoken BNC 2014 (14-25)) to investigate how Chinese learners differ from NNSs in their use of the DM you know and its functions in speech. The results show that there is a significant difference between the two corpora in terms of the frequency of use of you know. In terms of the functions of you know, the study shows that six functions can all be present in both corpora, although there are significant differences between the five functional dimensions, especially in introducing a claim linked to the prior discourse and highlighting particular points in the discourse. It is hoped to show empirically how Chinese learners and NSs use DMs differently.

Keywords: you know, discourse marker, native speaker, Chinese learner

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Authors: Isa Ahmadi, Ramin Khamedi

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In this paper, the elasto-plastic and cyclic torsion of a shaft is studied using a finite element method. The Prager kinematic hardening theory of plasticity with the Ramberg and Osgood stress-strain equation is used to evaluate the cyclic loading behavior of the shaft under the torsional loading. The material of shaft is assumed to follow the non-linear strain hardening property based on the Prager model. The finite element method with C1 continuity is developed and used for solution of the governing equations of the problem. The successive substitution iterative method is used to calculate the distribution of stresses and plastic strains in the shaft due to cyclic loads. The shear stress, effective stress, residual stress and elastic and plastic shear strain distribution are presented in the numerical results.

Keywords: cyclic loading, finite element analysis, Prager kinematic hardening model, torsion of shaft

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Authors: Swathi Gopakumar, Sruthi Krishna, Shivasubramani Krishnamoorthy

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Contactless, painless and radiation-free thermal imaging technology is one of the preferred screening modalities for detection of breast cancer. However, poor signal to noise ratio and the inexorable need to preserve edges defining cancer cells and normal cells, make the segmentation process difficult and hence unsuitable for computer-aided diagnosis of breast cancer. This paper presents key findings from a research conducted on the appraisal of two promising techniques, for the detection of breast cancer: (I) marker-controlled, Level-set segmentation of anisotropic diffusion filtered preprocessed image versus (II) Segmentation using marker-controlled level-set on a Gaussian-filtered image. Gaussian-filtering processes the image uniformly, whereas anisotropic filtering processes only in specific areas of a thermographic image. The pre-processed (Gaussian-filtered and anisotropic-filtered) images of breast samples were then applied for segmentation. The segmentation of breast starts with initial level-set function. In this study, marker refers to the position of the image to which initial level-set function is applied. The markers are generally placed on the left and right side of the breast, which may vary with the breast size. The proposed method was carried out on images from an online database with samples collected from women of varying breast characteristics. It was observed that the breast was able to be segmented out from the background by adjustment of the markers. From the results, it was observed that as a pre-processing technique, anisotropic filtering with level-set segmentation, preserved the edges more effectively than Gaussian filtering. Segmented image, by application of anisotropic filtering was found to be more suitable for feature extraction, enabling automated computer-aided diagnosis of breast cancer.

Keywords: anisotropic diffusion, breast, Gaussian, level-set, thermograms

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Authors: Sudheer Kumar Gundati, Umasankar Patro

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Carbon nanotube (CNT) – polymer composites have been extensively studied due to their exceptional electrical and mechanical properties. In the present study, poly(vinylidene fluoride) (PVDF) – multi-walled CNT composites were prepared by melt-blending technique using pristine (ufCNT) and a modified dilute nitric acid-treated CNTs (fCNT). Due to this dilute acid-treatment, the fCNTs were found to show significantly improved dispersion and retained their electrical property. The fCNT showed an electrical percolation threshold (PT) of 0.15 wt% in the PVDF matrix as against 0.35 wt% for ufCNT. The composites were made into films of thickness ~0.3 mm by compression-molding and the resulting composite films were subjected to various property evaluations. It was found that the water contact angle (WCA) of the films increased with CNT weight content in composites and the composite film surface became hydrophobic (e.g., WCA ~104° for 4 wt% ufCNT and 111.5° for 0.5 wt% fCNT composites) in nature; while the neat PVDF film showed hydrophilic behavior (WCA ~68°). Significant enhancements in the mechanical properties were observed upon CNT incorporation and there is a progressive increase in the tensile strength and modulus with increase in CNT weight fraction in composites. The composite films were tested for strain-sensing applications. For this, a simple and non-destructive method was developed to demonstrate the strain-sensing properties of the composites films. In this method, the change in electrical resistance was measured using a digital multimeter by applying bending strain by oscillation. It was found that by applying dynamic bending strain, there is a systematic change in resistance and the films showed piezo-resistive behavior. Due to the high flexibility of these composite films, the change in resistance was reversible and found to be marginally affected, when large number of tests were performed using a single specimen. It is interesting to note that the composites with CNT content notwithstanding their type near the percolation threshold (PT) showed better strain-sensing properties as compared to the composites with CNT contents well-above the PT. On account of the excellent combination of the various properties, the composite films offer a great promise as strain-sensors for structural health-monitoring.

Keywords: carbon nanotubes, electrical percolation threshold, mechanical properties, poly(vinylidene fluoride), strain-sensor, water contact angle

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Authors: H. A. Alguhi, W. A. Elsaigh

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This paper provides a tensile stress-strain (σ-ε) relationship for High-Strength Steel Fiber Reinforced Concrete (HSFRC). Load-deflection (P-δ) behavior of HSFRC beams tested under four-point flexural load were used with inverse analysis to calculate the tensile σ-ε relationship for various tested concrete grades (70 and 90MPa) containing 60 kg/m3 (0.76 %) of hook-end steel fibers. A first estimate of the tensile (σ-ε) relationship is obtained using RILEM TC 162-TDF and other methods available in literature, frequently used for determining tensile σ-ε relationship of Normal-Strength Concrete (NSC) Non-Linear Finite Element Analysis (NLFEA) package ABAQUS® is used to model the beam’s P-δ behavior. The results have shown that an element-size dependent tensile σ-ε relationship for HSFRC can be successfully generated and adopted for further analyzes involving HSFRC structures.

Keywords: tensile stress-strain, flexural response, high strength concrete, steel fibers, non-linear finite element analysis

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Authors: Priyashri Kamlesh Sridhar, Suranga Nanayakkara

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Current frameworks in assessment follow lesson delivery and rely heavily on test performance or teacher’s observations. This, however, neglects the underlying cognitive load during the learning process. Identifying the pivotal points when the load occurs helps design effective pedagogies and tools that respond to learners’ cognitive state. There has been limited research on quantifying cognitive load in preschoolers, real-time. In this study, we recorded electrodermal activity and heart rate variability (HRV) from 10 kindergarteners performing executive function tasks and Johnson Woodcock test of cognitive abilities. Preliminary findings suggest that there are indeed sensitive task-dependent markers in skin conductance (number of SCRs and average amplitude of SCRs) and HRV (mean heart rate and low frequency component) captured during the learning process.

Keywords: early childhood, learning, methodologies, pedagogies

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Authors: M. Azarbarmas, J. M. Cabrera, J. Calvo, M. Aghaie-Khafri

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The modeling of hot deformation behavior for unseen conditions is important in metal-forming. In this study, the hot deformation of IN718 has been characterized in the temperature range 950-1100 and strain rate range 0.001-0.1 s-1 using hot compression tests. All stress-strain curves showed the occurrence of dynamic recrystallization. These curves were implemented quantitatively in mathematics, and then constitutive equation indicating the relationship between the flow stress and hot deformation parameters was obtained successfully.

Keywords: compression test, constitutive equation, dynamic recrystallization, hot working

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Authors: Dipali Dhawan

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Cancerous epithelial cells are confined to a primary site by the continued expression of adhesion molecules and the intact basal lamina. However, as the cancer progresses some cells are believed to undergo an epithelial-mesenchymal transition (EMT) event, leading to increased motility, invasion and, ultimately, metastasis of the cells from the primary tumour to secondary sites within the body. These disseminated cancer cells need the ability to self-renew, as stem cells do, in order to establish and maintain a heterogeneous metastatic tumour mass. Identification of the specific subpopulation of cancer stem cells amenable to the process of metastasis is highly desirable. In this study, we have isolated and characterized cancer stem cells from luminal and basal breast cancer cell lines (MDA-MB-231, MDA-MB-453, MDA-MB-468, MCF7 and T47D) on the basis of cell surface markers CD44 and CD24; as well as Side Populations (SP) using Hoechst 33342 dye efflux. The isolated populations were analysed for epithelial and mesenchymal markers like E-cadherin, N-cadherin, Sfrp1 and Vimentin by Western blotting and Immunocytochemistry. MDA-MB-231 cell lines contain a major population of CD44+CD24- cells whereas MCF7, T47D and MDA-MB-231 cell lines show a side population. We observed higher expression of N-cadherin in MCF-7 SP cells as compared to MCF-7NSP (Non-side population) cells suggesting that the SP cells are mesenchymal like cells and hence express increased N-cadherin with stem cell-like properties. There was an expression of Sfrp1 in the MCF7- NSP cells as compared to no expression in MCF7-SP cells, which suggests that the Wnt pathway is expressed in the MCF7-SP cells. The mesenchymal marker Vimentin was expressed only in MDA-MB-231 cells. Hence, understanding the breast cancer heterogeneity would enable a better understanding of the disease progression and therapeutic targeting.

Keywords: cancer stem cells, epithelial to mesenchymal transition, biomarkers, breast cancer

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Authors: Hayat Omar

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Languages provide speakers with a wide range of linguistic units to organize and deliver information. There are several ways to verbally express the mental representations of events. According to the linguistic tools they have acquired, speakers select the one that brings out the most communicative effect to convey their message. Our study focuses on two markers, -mm and dämmo, in Amharic (Ethiopian Semitic language). Our aim is to examine, from a developmental perspective, how they are used by speakers. We seek to distinguish the communicative and pragmatic functions indicated by means of these markers. To do so, we created a corpus of sixty narrative productions of children from 5-6, 7-8 to 10-12 years old and adult Amharic speakers. The experimental material we used to collect our data is a series of pictures without text 'Frog, Where are you?'. Although -mm and dämmo are each used in specific contexts, they are sometimes analyzed as being interchangeable. The suffix -mm is complex and multifunctional. It marks the end of the negative verbal structure, it is found in the relative structure of the imperfect, it creates new words such as adverbials or pronouns, it also serves to coordinate words, sentences and to mark the link between macro-propositions within a larger textual unit. -mm was analyzed as marker of insistence, topic shift marker, element of concatenation, contrastive focus marker, 'bisyndetic' coordinator. On the other hand, dämmo has limited function and did not attract the attention of many authors. The only approach we could find analyzes it in terms of 'monosyndetic' coordinator. The paralleling of these two elements made it possible to understand their distinctive functions and refine their description. When it comes to marking a referent, the choice of -mm or dämmo is not neutral, depending on whether the tagged argument is newly introduced, maintained, promoted or reintroduced. The presence of these morphemes explains the inter-phrastic link. The information is seized by anaphora or presupposition: -mm goes upstream while dämmo arrows downstream, the latter requires new information. The speaker uses -mm or dämmo according to what he assumes to be known to his interlocutors. The results show that -mm and dämmo, although all the speakers use them both, do not always have the same scope according to the speaker and vary according to the age. dämmo is mainly used to mark a contrastive topic to signal the concomitance of events. It is more commonly used in young children’s narratives (F(3,56) = 3,82, p < .01). Some values of -mm (additive) are acquired very early while others are rather late and increase with age (F(3,56) = 3,2, p < .03). The difficulty is due not only because of its synthetic structure but primarily because it is multi-purpose and requires a memory work. It highlights the constituent on which it operates to clarify how the message should be interpreted.

Keywords: acquisition, cohesion, connection, contrastive topic, contrastive focus, discourse marker, pragmatics

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Authors: M. Ghosh, A. Miroux, L. A. I. Kestens

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At molecular or micrometre scale, practically all materials are neither homogeneous nor isotropic. The concept of texture is used to identify the structural features that cause the properties of a material to be anisotropic. For metallic materials, the anisotropy of the mechanical behaviour originates from the crystallographic nature of plastic deformation, and is therefore controlled by the crystallographic texture. Anisotropy in mechanical properties often constitutes a disadvantage in the application of materials, as it is often illustrated by the earing phenomena during drawing. However, advantages may also be attained when considering other properties (e.g. optimization of magnetic behaviour to a specific direction) by controlling texture through thermo-mechanical processing). Nevertheless, in order to have better control over the final properties it is essential to relate texture with materials processing route and subsequently optimise their performance. However, up to date, few studies have been reported about the evolution of texture in 6061 aluminium alloy during warm processing (from room temperature to 250ºC). In present investigation, recrystallized 6061 aluminium alloy samples were subjected to tensile and plane strain compression (PSC) at room and warm temperatures. The gradual change of texture following both deformation modes were measured and discussed. Tensile tests demonstrate the mechanism at low strain while PSC does the same at high strain and eventually simulate the condition of rolling. Cube dominated texture of the initial rolled and recrystallized AA6061 sheets were replaced by domination of S and R components after PSC at room temperature, warm temperature (250ºC) though did not reflect any noticeable deviation from room temperature observation. It was also noticed that temperature has no significant effect on the evolution of grain morphology during PSC. The band contrast map revealed that after 30% deformation the substructure inside the grain is mainly made of series of parallel bands. A tendency for decrease of Cube and increase of Goss was noticed after tensile deformation compared to as-received material. Like PSC, texture does not change after deformation at warm temperature though. n-fibre was noticed for all the three textures from Goss to Cube.

Keywords: AA 6061, deformation, temperature, tensile, PSC, texture

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Authors: Jakub Bartus, Jaroslav Odrobinak

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In general, composite steel and concrete structures present an effective structural solution utilizing the full potential of both materials. As they have numerous advantages on the construction side, they can greatly reduce the overall cost of construction, which has been the main objective of the last decade, highlighted by the current economic and social crisis. The study represents not only an analysis of composite beams’ behavior having web openings but emphasizes the influence of these openings on the total strain distribution at the level of the steel bottom flange as well. The major investigation was focused on a change in structural performance with respect to various layouts of openings. Examining this structural modification, an improvement of load carrying capacity of composite beams was a prime objective. The study is divided into analytical and numerical parts. The analytical part served as an initial step into the design process of composite beam samples, in which optimal dimensions and specific levels of utilization in individual stress states were taken into account. The numerical part covered the discretization of the preset structural issue in the form of a finite element (FE) model using beam and shell elements accounting for material non–linearities. As an outcome, several conclusions were drawn describing and explaining the effect of web opening presence on the structural performance of composite beams.

Keywords: beam, steel flange, total strain, web opening

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Authors: Mark Bezner, Shani Deri, Tom Trigano, Kfir Ben-Harush

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Intermediate filament (IF) proteins-based fibers are among the toughest fibers in nature, as was shown by native hagfish slime threads and by synthetic fibers that are based on type V IF-proteins, the nuclear lamins. It is assumed that their mechanical performance stems from two major factors: (1) the transition from elastic -helices to stiff-sheets during tensile load; and (2) the specific organization of the coiled-coil proteins into a hierarchical network of nano-filaments. Here, we investigated the interrelationship between these two factors by using wet-spun fibers based on C. elegans (Ce) lamin. We found that Ce-lamin fibers, whether assembled in aqueous or alcoholic solutions, had the same nonlinear mechanical behavior, with the elastic region ending at ~5%. The pattern of the transition was, however, different: the ratio between -helices and -sheets/random coils was relatively constant until a 20% strain for fibers assembled in an aqueous solution, whereas for fibers assembled in 70% ethanol, the transition ended at a 6% strain. This structural phenomenon in alcoholic solution probably occurred through the transition between compacted and extended conformation of the random coil, and not between -helix and -sheets, as cycle analyses had suggested. The different transition pattern can also be explained by the different higher order organization of Ce-lamins in aqueous or alcoholic solutions, as demonstrated by introducing a point mutation in conserved residue in Ce-lamin gene that alter the structure of the Ce-lamins’ nano-fibrils. In addition, biomimicking the layered structure of silk and hair fibers by coating the Ce-lamin fiber with a hydrophobic layer enhanced fiber toughness and lead to a reversible transition between -helix and the extended conformation. This work suggests that different hierarchical structures, which are formed by specific assembly conditions, lead to diverse secondary structure transitions patterns, which in turn affect the fibers’ mechanical properties.

Keywords: protein-based fibers, intermediate filaments (IF) assembly, toughness, structure-property relationships

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